Differences in neutralizing antigenicity between laboratory and clinical isolates of HCoV-229E isolated in Japan in 2004–2008 depend on the S1 region sequence of the spike protein

Abstract

Human coronavirus (HCoV) is a causative agent of the common cold. Although HCoV is highly prevalent in the world, studies of the genomic and antigenic details of circulating HCoV strains have been limited. In this study, we compared four Japanese isolates with the standard HCoV-229E strain obtained from ATCC (ATCC-VR740) by focusing on the spike (S) protein, a major determinant of neutralizing antigen and pathogenicity. The isolates were found to have nucleotide deletions and a number of sequence differences in the S1 region of the S protein. We compared two of the Japanese isolates with the ATCC-VR740 strain by using virus-neutralizing assays consisting of infectious HCoV-229E particles and vesicular stomatitis virus (VSV)-pseudotyped virus carrying the HCoV-229E S protein. The two clinical isolates (Sendai-H/1121/04 and Niigata/01/08) did not react with antiserum to the ATCC-VR740 strain via the neutralizing test. We then constructed a pseudotype VSV-harboured chimeric S protein with the ATCC S1 and Sendai S2 regions or that with Sendai S1 and ATCC S2 regions and compared them by a neutralization test. The results revealed that the difference in the neutralizing antigenicity depends on the S1 region. This different antigenic phenotype was also confirmed by a neutralizing test with clinically isolated human sera. These results suggest that the HCoV-229E viruses prevalent in Japan are quite different from the laboratory strain ATCC-VR740 in terms of the S sequence and neutralization antigenicity, which is attributed to the difference in the S1 region. INTRODUCTIONThe coronavirus (CoV) is an enveloped virus with a singlestranded, positive RNA genome of about 30 kb. Moreover, the virion is approximately 80-120 nm in diameter (Lai, 1990;Lai & Cavanagh, 1997) and is characterized in electron micrographs by its crown-like appearance of spikes protruding from the virion envelope (Lai, 1990;Lai & Cavanagh, 1997). A variety of virus species that infect domestic animals, as well as pets, are classified as CoVs. Some CoVs are highly pathogenic while others exhibit low virulence. Before severe acute respiratory syndrome (SARS)-CoV was reported in 2003(Peiris et al., 2004, only low-virulence human CoVs (HCoVs), such as 229E or OC43, were known to exist. After the SARS outbreak, more virulent HCoVs, such as HKU1 (Woo et al., 2005) and NL63 (van der Hoek et al., 2004), were isolated from humans with respiratory diseases.HCoV-229E, which was first reported in 1966 (Hamre & Procknow, 1966), is a prototype of HCoV and belongs to the genus Alphacoronavirus, together with HCoV-NL63. Both cause respiratory diseases in human infants (van der Hoek et al., 2004). Dijkman et al. (2008) reported that 75 and 65 % of the children between the age of 2.5 and 3.5 years were seropositive for NL63 and HCoV-229E, respectively, and most of the children seroconverted against both viruses by the age of six. This may mean that most people experience acute infection by these HCoVs during their childhood. However, while HCoV is highly A supplementar...

Inhibitory effects of glycopyrronium, formoterol, and budesonide on coronavirus HCoV-229E replication and cytokine production by primary cultures of human nasal and tracheal epithelial cells

“…A stock of clinically isolated HCoV-229E [2] was prepared by infecting Rhesus monkey kidney epithelial (LLC-MK2) cells.…”

“…Human coronaviruses (HCoV)-229E and HCoV-OC43 cause the common cold [1,2] and exacerbate chronic obstructive pulmonary disease (COPD) and bronchial asthma [3,4]. HCoV-229E binds to the aminopeptidase N receptor (CD13) [5] and enters cells via endosomal and cell-surface pathways using proteases [6,7].…”

Development of fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) using quenching probes for the detection of the Middle East respiratory syndrome coronavirus

“…Human metapneumovirus (HMPV; Sendai-H/2404/2003) was obtained from the Virus Research Center, Sendai Medical Center, Japan. Human coronavirus (HCoV)-229E isolates ATCC VR-740 and Sendai-H/1121/04 (Shirato et al, 2012) were used. HCoV-NL63 was supplied by Dr. Lia van der Hoek, University of Amsterdam, the Netherlands.…”

HeLa-ACE2-TMPRSS2 Cells Are Useful for the Isolation of Human Coronavirus 229E

“…In general, the primary isolation of HCoV-229E is difficult, and HCoV-229E strains cannot be grown easily in cell cultures without extensive adaptation by passaging (4). Secondary human embryonic kidney cells, human embryonic intestine MA-177 cells, the monkey kidney epithelial cell LLC-MK2 cell line, human embryonic fibroblasts, and the human colon epithelial cancer CaCo-2 cell line have been reported as permissive cells for the isolation of HCoV-229E from clinical specimens (5)(6)(7). In addition, we have reported the isolation of HCoV-229E on rhabdomyosarcoma RD-18S cells by chance, which, when infected, showed picornavirus-like cytopathic effects (CPEs) (8).…”